Unhealthy C2C12 myoblasts - (Apr/20/2006 )

I am worried about my C2C12 myoblasts at the moment - they were looking "spindley" last week so I got rid of them, made up fresh media and fresh PBS and broke out a fresh batch of cells. However they still looked the same today (please see picture attached). Should I be worried? I am maintaining them in DMEM (Sigma #D6429) supplemented with 20%FBS, 2mM glutamine and Pen/Strep. I use trypsin to lift them. I am just sub-culturing them at the moment and am not trying to differentiate them. I'm also maintaining HeLa cells at the moment and they look fine.

Thanks for you attention.

are the two types of cells in the same incubator? check the water pan, and confirm the CO2 percentage....

could there have been a problem with the procedure when your myoblasts were frozen? and, last, have you checked for mycoplasma?

Actually, I regard that your C2C12 has been differentiated into others cell ( some are neural phenotype and others are myotube). I has also cultured the C2C12 months ago and found that it has the ability differentiated into neural cell and myotube under low serum(DMEM w/2% horse serum). They always formed mixed population in normal conditions and don't forget not to let them become confluence in the culture. Otherwise , they may differenitate into others....